My research projects under Suzanne Michalek, Professor of Microbiology have been involved with the characterization of inflammatory cytokines in gingival crevicular fluid (GCF) and analysis of messenger RNA (mRNA) for local cytokine production in adult periodontitis. My initial work with the GCF will be presented at the 1992 American Society for Microbiology Annual Session, and the mRNA work will be presented at the 1992 International Association for Dental Research Annual Session. I have been studying the presence of the cytokines interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-a), and transforming growth factor-beta (TGF-b) in the GCF of patients with advanced periodontal disease. Crevicular fluid samples were taken from sites with attachment loss and as well as healthy controls. The GCF samples were analysed by SDS-PAGE, transferred to nitrocellulose membranes, and probed with cytokine specific antibodies. We demonstrated the presence of these cytokines by developing the blots with an alkaline phosphatase procedure. The local production of the cytokines interleukins-1 beta (IL-1b), interleukin-1 alpha (IL-la), TNF-a, TGF-b, IL-6, and interleukin-8 (IL-8) by the gingival mononuclear cell (GMNC) is also being studied in a project involving the isolation and purification of mRNA from the GMNC of periodontally inflamed tissues. Even very small amounts of mRNA can then be reverse transcribed into cDNA amplified by the RT-PCR and method. Our studies thus far indicate the presence of message for IL-1b, TNF-a, IL-6, and Il-8 in all samples tested, while IL-1a and TGF-b were only found in 50% of the samples. My future research plans include the use of bioassay techniques for the presence and amounts of active cytokines in the GCF project, quantitative PCR methods involving cotranscription of "housekeeping" genes, and in situ hybridization studies for localization of the cells responsible for the production of these cytokines in periodontally inflamed tissues.